The use of confocal microscopy establishes methodology to count the keratocytes in the corneal stroma, and measure the density of the ADAScs in the implanted tissue, as well as in the corneal stroma.
After three months, it became recellularized by the patients’ own keratocytes.
“There was a migration of keratocytes from the host stroma toward the lamina,” said Dr. Zarif. The posterior surface of the lamina recellularized before the anterior surface. After a year, the keratocytes populated the lamina in a similar way to a normal cornea stroma.
The third group, implanted with recellularized lamina, showed structures like normal keratocytes after one month. These were more voluminous in the posterior surface of the lamina. With some patients they had dendritic shape after one year of implantation.
In all three groups, 12 months after the surgery the researcher noted gradual and significant increase in the cellularity of the anterior and posterior stromas of the patients in comparison to the preoperative density, with a p value for this difference of less than 0.001 in the anterior and posterior stroma.
Mid corneal stroma
As the study progressed, the research team continued to make new discoveries in each of the groups.
In the first group, the researchers noted a gradual but significant increase of the cellularity of the mid corneal stroma (p < 0.001).
In groups 2 and 3, one year after surgery, the researchers observed a significant increase (p < 0.001) in cellularity in the anterior and posterior surfaces and within the lamina.
They found that cell density was significantly higher in patients implanted with recellularized laminas than in those implanted with decellularized laminas, both on the anterior surface and within the lamina (p =0.011), and on the posterior surface of the lamina (p = 0.029).
After making its initial findings on cell density, the research team continued to follow up on this measurement.
After one year, group 3 had greater cell density than either group 2 or group 1 in the anterior stroma. It had greater cell density than group 2 in the anterior stroma, the anterior surface of the lamina, the mid stroma of the lamina and the posterior surface of the lamina.
In conclusion, Dr. El Zarif said, confocal microscopy is an essential tool for the evaluation and monitoring “in vivo” of the ADASC.
“It allowed a qualitative and quantitative assessment of the cells through the experiment,” she said. “And it allowed monitoring of the progressive morphological changes that occurred in the implanted tissue in decellularized and recellularized laminas.”
Dr. El Zarif added that the technique assisted in determining the change in the cells’ densities in the grafted tissue as well as in all the posterior and anterior corneal stroma.
Mona El Zarif, OD, Msc
E: [email protected]
Dr. El Zarif has no financial disclosures.